LOCUS pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP + Sense IRES 8011 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses a GFP circular RNA containing the sense EMCV IRES in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP + Sense IRES
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8011)
AUTHORS Kramer MC, Liang D, Tatomer DC, Gold B, March ZM, Cherry S, Wilusz
JE
TITLE Combinatorial control of Drosophila circular RNA expression by
intronic repeats, hnRNPs, and SR proteins.
JOURNAL Genes Dev. 2015 Oct 8.
PUBMED 26450910
REFERENCE 2 (bases 1 to 8011)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8011
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1656..2229
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 1679..2231
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1823..1840)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2050..2069
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind complement(3605..3622)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3611..3835
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3881..4309
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3968..3987)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4178..4199
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4318..4338)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4323..4652
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4503..4638
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4565..4584
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4719..5513
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4773..4792)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5383..5402
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5687..5808
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5724..5743)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5778..5797
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5857..5873)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5857..5873)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5870..5892)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5881..5897
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5905..5935)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5950..5971
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6088..6105)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6259..6844)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6339..6358)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7015..7875)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7638..7657
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7876..7980)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcagt
961 gacagtggag attgtacagt tttttcctcg atttgtcagg attttttttt tttgacggag
1021 tttaacttct tgtctcccag gtaggaagtg cagtggcgta atctcggctc actacaacct
1081 ccacctcctg ggttcaagcg tttctcctgc ctcagctttc cgagtagctg ggattacagg
1141 cgcctgccac catgccctgc tgacttttgt atttttagta gagacggggt ttcaccatgt
1201 tggccaggct ggtcttgaac tcctgaccgc aggcgattgg cctgcctcgg cctcccaaag
1261 tgctgagatt acaggcgtga gccaccaccc ccggcctcag gagcgttctg atagtgcctc
1321 gatgtgctgc ctcctataaa gtgttagcag cacagatcac tttttgtaaa ggtacgtact
1381 aatgactttt tttttatact tcaggatggc agcgtgcagc tggccgacca ctaccagcag
1441 aataccccca tcggcgatgg ccctgtgctg ctgcccgata accactacct gtccacccag
1501 agcgccctgt ccaaggaccc caacgagaag cgcgatcaca tgatctactt cggcttcgtg
1561 accgccgccg gcatcaccca cggcatggac gagctgtaca agtgaatcga taccggttta
1621 attaacacgt gggtaaccgt tccgagtcgt taacgcccct ctccctcccc cccccctaac
1681 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
1741 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
1801 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
1861 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
1921 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
1981 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
2041 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
2101 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
2161 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
2221 cgatgataat atggcgttaa ccaccatggc cagcaagggc gccgagctgt tcaccggcat
2281 cgtgcccatc ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg
2341 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
2401 caagctgcct gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt
2461 ctcacgctac cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg
2521 ctacatccag gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga
2581 ggtgaagttc gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa
2641 ggaggatggc aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta
2701 catcatgacc gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat
2761 cgaggtaaga agcaaggttt catttagggg aagggaaatg attcaggacg agagtctttg
2821 tgctgctgag tgcctgtgat gaagaagcat gttagtcctg ggcaacgtag cgagacccca
2881 tctctacaaa aaatagaaaa attagccagg tatagtggcg cacacctgtg attccagcta
2941 cgcaggaggc tgaggtggga ggattgcttg agcccaggag gttgaggctg cagtgagctg
3001 taatcatgcc actactccaa cctgggcaac acagcaagga ccctgtctca aaagctactt
3061 acagaaaaga attaggctcg gcacggtagc tcacacctgt aatcccagca ctttgggagg
3121 ctgaggcggg cagatcactt gaggtcagga gtttgagacc agcctggcca acatggtgaa
3181 accttgtctc tactaaaaat atgaaaatta gccaggcatg gtggcacatt cctgtaatcc
3241 cagctactcg ggaggctgag gcaggagaat cacttgaacc caggaggtgg aggttgcagt
3301 aagccgagat cgtaccactg tgctctagcc ttggtgacag agcgagactg tcttaaaaaa
3361 aaaaaaaaaa aaaaaagaat taattaaaaa tttaaaaaaa aatgaaaaaa agctgcatgc
3421 ttgttttttg tttttagtta ttctacattg ttgtcattat taccaaatat tggggaaaat
3481 acaacttaca gaccaatctc aggagttaaa tgttactacg aaggcaaatg aactatgcgt
3541 aatgaacctg gtaggcatta gcggccgctc gagtctagag ggcccgttta aacccgctga
3601 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
3661 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
3721 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
3781 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
3841 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
3901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
3961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
4021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
4081 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
4141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
4201 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
4261 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
4321 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
4381 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
4441 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
4501 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
4561 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
4621 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
4681 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
4741 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
4801 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
4861 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
4921 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
4981 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
5041 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
5101 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
5161 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
5221 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
5281 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
5341 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
5401 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
5461 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
5521 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
5701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
5761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
5821 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
5881 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
5941 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
6001 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
6061 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
6121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
6181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
6241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
6301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
6361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
6421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
6481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
6541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
6601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
6661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
6721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
6781 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
6841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
6901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
6961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
7021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
7081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
7141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
7201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
7261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
7321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
7381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
7441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
7501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
7561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
7621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
7681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
7741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
7801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
7861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
7921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
7981 cacatttccc cgaaaagtgc cacctgacgt c
//